OPTICALLY ENHANCED OLIGONUCLEOTIDES TO SCREEN FOR METABOLIC DISEASES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 目前的代谢性疾病筛查方法过于昂贵，昂贵且耗时，无法广泛测试所有新生儿。因此，许多代谢性疾病多年未被发现和治疗，并且仅对一种最常见的代谢性疾病进行筛查。使用所提出的溶胶-凝胶支持的核酸适体-染料混合和测量测定法在尿液中允许成本有效的大规模筛选。 该研究致力于开发新的，高选择性和灵敏的分子传感器，在类固醇代谢物存在下改变颜色或荧光特性。几种类固醇是先天性代谢疾病的疾病标志物。我们感兴趣的是映射三路路口的基于阿托伐他汀的粘合剂（适体），以确定一般原则，可用于了解分子识别的荧光染料，可见染料，类固醇，及其代谢物的适体。 适体-染料复合物的筛选将通过产生适合于大规模筛选和监测类固醇或由错误的类固醇代谢引起的代谢物的具有成本效益的、快速的、混合和测量的方法来解决重要的医学需求。使用组合96孔板方法将发现与结合适体和代谢物形成强复合物的有机染料。我们的小组已经建立了可靠的紫外-可见和荧光光谱法的适体结合测定。 此外，适体将被纳入胶体硅酸盐聚合物的手持固体传感器设备的发展，用于医疗应用。“溶胶-凝胶”基于固体胶体硅酸盐，其将形成多孔硬凝胶并将适体-染料复合物掺入凝胶内，从而允许将新的分子传感器附着到固体支持物中。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Current screening methods for metabolic diseases are too costly, expensive, and time-consuming for broadly testing all newborns. Thus, many metabolic diseases go undetected and untreated for many years and screening is implemented only for the one most common metabolic disease. Using the proposed sol-gel supported aptamer-dye mix-and-measure assays in urine allows for cost-effective mass screening. The roposed research is dedicated to the development of new, highly selective, and sensitive molecular sensors that change color or fluorescence properties in the presence of steroid metabolites. Several steroids are disease markers for inborn metabolic diseases. We are interested in mapping three-way junctions of oligonucleotide-based binders (aptamers) to determine general principles that can be used in understanding molecular recognition of fluorescent dyes, visible dyes, steroids, and their metabolites by aptamers. The screening of aptamer-dye complexes will address an important medical need by generating cost-effective, rapid, mix-and-measure methods suitable for mass screening and monitoring of steroids or metabolites caused by faulty steroid metabolism. Organic dyes that form strong complexes with binding aptamers and metabolites will be found using a combinatorial 96-well plate approach. Reliable aptamer binding assays by UV-Vis and fluorescence spectroscopy have been established by our group. Furthermore, aptamers will be incorporated into colloidal silicate polymers for the development of handheld solid sensor devices to be used in medicinal applications. "Sol-gels" are based on solid colloidal silicates that will form a porous hard gel and will incorporate the aptamer-dye complexes within the gel, allowing for attaching the new molecular sensors into a solid support.